Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (32): 5953-5958.doi: 10.3969/j.issn.2095-4344.2012.32.012

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Osteogenic and chondrogenic differentiation potential of human adipose-derived mesenchymal stem cells

Zhu Yin1, Xu Wei-yuan2, Wang Wen-jia3, Zhang Xing-xiang2, Yan Fei2, Sha Wei-ping2, Zhu Xian-wei1   

  1. 1Clinical Medical College of Jiangsu University, Zhenjiang 212013, Jiangsu Province, China;
    2Department of Orthopedics, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China;
    3Central Laboratory, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China
  • Received:2012-04-25 Revised:2012-06-11 Online:2012-08-05 Published:2012-08-05
  • Contact: Xu Wei-yuan, M.D., Professor, Chief physician, Department of Orthopedics, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China xwyl43@163.com
  • About author:Zhu Yin★, Studying for master's degree, Clinical Medical College of Jiangsu University, Zhangjiagang 215600, Jiangsu Province, China zhuyin871113@qq.com

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Abstract:

BACKGROUND: Human adipose-derived mesenchymal stem cells are a kind of adult stem cells with the high ability of proliferation and multi-lineage differentiation. They can be acquired from the cosmetic liposuction operation with rich sources of raw material and the selections are very convenient. They have a potential value in the field of bioremediation.
OBJECTIVE: To establish a method of isolating and culturing human adipose-derived mesenchymal stem cells and investigate the proficiency of basic biological characteristics and the potential ability of osteogenic and chondrogenic differentiation.
METHODS: The adipose tissue was taken from the liposuction operation, human adipose-derived mesenchymal stem cells were isolated by type II collagen enzyme digestion and then cultured in vitro. Cell morphology was observed, cell cycle was determined, and cell surface markers were identified with flow cytometry. Passage 3 cells were assigned to two induction groups A, B and two control groups A, B. Then the cells were induced to differentiate into osteoblasts and chondrocytes with different culture media in vitro. The differentiated cells were identified by histochemical staining, immunocytochemical staining and RT-PCR.
RESULTS AND CONCLUSION: After in vitro culture, human adipose-derived mesenchymal stem cells showed the desmoids shape. The primary cells adhered to the wall in 24 hours, and then formed the colony after 5-7 days. The passaged cells adhered to the wall in 4-6 hours and maintained the same shape. Cell cycle studies showed that cells at G0/G1, S and G2/M accounted for (88±2)% (88±2)% and 0.03% respectively. The flow cytometry examination showed that the cells were positive for CD29 and CD105 but they were negative for CD45. RT-PCR showed that after osteogenic induction, human adipose-derived mesenchymal stem cells had positive osteopontin mRNA expression; after chondrogenic induction, cells had positive type II collagen mRNA expression. Human adipose-derived mesenchymal stem cells were successfully isolated and cultured and the cells exhibit the potential to differentiate into osteoblasts and chondrocytes.

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